6-methyl, delta1, 4, 3 keto androstene derivatives



6-METHYL, A 3 KETU ANDROSTENE DERHVATKVES Jean P. Rosseiet, Kalamazoo,Mich, assignor to The UpjohnCompany, Kalamazoo, Mich, a corporation ofMichigan No Drawing. Application December 11, 1957 Serial No. 701,986

7 Claims. (Cl. 260-37.3)

The present invention relates to steroid compounds and is particularlyconcerned with 1-dehydro-6-methyltestosterone and the 17-esters thereofand l-dehydro-6-methyladrenosterone.

The new compounds and the process of the present invention areillustratively represented by the following formulae:

wherein R is selected from the suhstituents consisting of keto oxygen;

and

H AcO wherein Ac is the acyl radical of a hydrocarbon carboxylic acidcontaining from one to twelve carbon atoms, inclusive, and R is selectedfrom the group consisting of 6ex-methyl and 6fl-methyl.

The process of the instant invention comprises submitting6-methyl-ll-ketoprogesterone (I) to the fermentative action of anorganism of a species of the genus Septomyxa to give simultaneouslyl-dehydro-6-methyladrenosterone andl-dehydro-6-methyl-1l-ketotestosterone (Illa). From1-dehydro-6-methyl-1l-ketotestosterone by oxidation with chromic acidthe l-dehydro-6- methyladrenosterone (II) can be obtained. Byesterifying in conventional mannerl-dehydro-6-methy1-ll-ketotestosterone with acid anhydrides, acidhalides, such as an acyl chloride, acyl bromide; or ketenes andisopropenylacylates, the corresponding l7-ester of l-dehydro-6-methy1-ll-ketotestosterone'(III/J) is obtained. Details of thesereactions are shown in the examples.

It is an object of the instant invention to provide 1-dehydro-6-methyladrenosterone, 1-dehydro-6-methyl-l1- ketotestosteroneand the 17-esters thereof as well as methods for the productiontherefor, especially in the n-epimeric form, i. e.l-dehydro-6a-methyladrenosterone and l-dehydro-6a-methyl-ll-ketotestosterone and the 17- esters thereof.

The new compounds 1-dehydro-6-methyladrenosterone, and1-dehydro-6-methyl-ll-ketotestosterones and the 17- ice estersespecially as 6a-methyl epimers thereof have gonadotropin inhibition andparticularly inhibit the follicle stimulating hormone (FSH). Thecompound is therefore useful in preventing estrus in dogs and othermammals, as oral contraceptives, or in the treatment of nymphomania,dysmenorrhea or any imbalance between the follicle stimulating hormoneand the luteinising hormone.

The following examples are illustrative of the process and product ofthe present invention, but are not to be construed as limiting.

EXAMPLE 1 1 -dehydro-6a-methyladrenosterone and 1 -dehydr0-6 xmethyl-1]-ket0test0ster0ne Ten liters of a medium consisting of one percentCerelose dextrose, two percent corn steep liquor containing sixtypercent solids, was adjusted to pH 4.85 with sodium hydroxide. Tenmilliliters of lard oil containing 0.1 to 2.0 percent octadecanol wasadded to prevent foaming. The medium was steam sterilized. Upon cooling,the sterile medium was inoculated with 500 milliliters of a nineteenhour growth from spores of Septomyxa afiinz's ATCC 6737. The medium wasagitated (300 R. P. M.) and sparged with sterile air at the rate of 0.1liter of air per minute. After culturing for 24 hoursthere was addedfour grams of 6m-methyl-ll-ketoprogesterone [G. B. Spero et al., I. Am.Chem. Soc. 78, 6213 (1956)], dissolved in 35 milliliters of acetone.Fermentation of the substrate was maintained for 24 hours at which timethe pH was 7.15. The fermentation broth was then filtered to separatethe mycelium. The mycelium was washed once with one liter of acetone andthen twice with one-liter portions of methylene chloride. The acetoneand methylene chloride washings were combined with an additional fourliters of methylene chloride and this seven liters of extract andsolvent was then used to extract the filtered beer. Upon separation ofthe extract from the beer, the beer was twice more extracted withthree-liter volumes of methylene chloride. All of the acetone andmethylene chloride extracts were combined and washed with 1200milliliters of two percent sodium bicarbonate solution, and then with1200 milliliters of water. The washed solvent extract was then driedwith anhydrous sodium sulfate and vacuum-evaporated to leave 5.2 gramsof dry extracts.

The extracts were dissolved in milliliters of benzene and fractionatedover a column of 200 grams of alumina (acid washed, dried at 120 degreescentigrade) using 120 milliliter portions of developing solvent as shownin Table I.

TABLE I Residue in Milligrams benzene discarded benzenezether 3:1 1

.do ethegchloroform 19: 1

ether-chloroform 3:1..

d0 a. methanol Fractions four, five and six were combined andrecrystallized from hot benzene to give 353 milligrams ofl-dehydro-6a-methyladrenosterone of melting point 274 to 276 degreesCentigrade with decomposition and rotation [@1 of plus 230 degrees inchloroform.

Analysis.-Calcd. for 1-1 0 C, 76.89; H, 7.74. Found: C, 76.67; H, 7.94.

Thereafter fractions nine through fourteen were combined and twicerecrystallized from methanol chloride ether to give 980 milligrams ofcrystals of l-dehydro- 6u-methyl-1l-ketotestosterone of melting point209 to 210 degrees centigrade and rotation [al of plus 148 degrees inchloroform.

Analysis.-Calcd. for (1 1-1 0 C, 76.40; H, 8.34. Found: C, 76.42; H.8.30.

In the same manner, using ofi-rnethyl-l l-ketoprogesterone as startingmaterial, the above fermentation is pro ductive of a mixture of1-dehydro-6B-methyladrenosterone and1-dehydro-6fi-methyl-ll-ketotestosterone.

EXAMPLE 2 1 -dehydr0-6oc-methyladrenosterone from I-dehydr0-6mmethy [-11 -ke totestosterone To 100 milligrams ofl-dehydro-Ga-methyl-ll-ketotestosterone, dissolved in glacial aceticacid, was added forty milligrams of chromium oxide in two milliliters ofglacial acetic acid. The mixture was kept at room temperature forsixteen hours, then diluted with four milliliters of methanol. After twohours at room temperature, the mixture was evaporated to dryness invacuo, the residue thus obtained dissolved in methylene chloride,decolorized with carbon, filtered and evaporated to dryness. Theresidue, 89 milligrams, gave upon crystallization from acetone-ether 64milligrams of colorless needles of melting point 264 to 270 degreescentigrade. This melting point was not depressed when the substance wasmixed with l.-dehydro-6sz-rnethyladrenosterone, as obtained from thefermentation in Example 1.

EXAMPLE 3 I-dehydro-6a-methyl-1I-ketotestosterone 17-acetate A mixturewas prepared containing in one milliliter of pyridine 100 milligrams ofl-dehydro-rx-methyl-llketotestosterone. To this solution was added onemilliliter of acetic anhydride. The mixture was allowed to stand at roomtemperature for a period of four hours and was thereupon diluted withtwenty milliliters of water. The aqueous mixture was then cooledovernight to about zero to five degrees centigrade and filteredthereafter. The precipitate obtained from the filtration was twicerecrystallized from methylene chloride and ether to givel-dehydro-6ot-methyl-ll-ketotestosterone 17-acetate.

EXAMPLE 4 J -dehydr0-6u-methyl-11-ketotest0ster0ne 1 7-benzoate Amixture was prepared containing in one milliliter of pyridine and onemilliliter of benzoyl chloride 0.1 gram ofl-dehydro-6m-methyl-ll-ketotestosterone. This mixture was allowed tostand for eighteen hours overnight, was thereupon diluted with fiftymilliliters of water and refrigerated to about five degrees centigrade.Thereafter the mixture was extracted with two 25-milliliter portions ofmethylene chloride, the methylene chloride extracts were washed twicewith water, dried over anhydrous sodium sulfate and evaporated. Thesolid was three times recrystallized from methanol to givel-dehydro-61-methyl-1l-ketotestosterone 17-benzoate.

EXAMPLE 1 -dehydr0-6a--melhy [-1 1 -ket0test0ster0ne 1 7-propi0nate Inthe same manner as shown in Example 3, treating1-dehydro-6m-methyl-1l-ketotestosterone at room temperature in pyridinesolution with propionic anhydride 4 results in1-dehydro-6a-methyl-ll-ketotestosterone 17- propionate, a light coloredcrystalline solid.

EXAMPLE 6 1 -dehydro-6ot-methyl-1 1 -ket0 testosterone 17-laurate In thesame manner as shown in Example 3, treating1-dehydro-6a-methyl-ll-ketotestosterone at room temperature in pyridinesolution with lauric anhydride results inl-dehydro-6u-methyl-ll-ketotestosterone 17- laurate.

EXAMPLE 7 1 -dehydr0-6a-methyl-1 1 -ket0test0ster0ne 17-trimethylacetate In the same manner as shown in Example 3, treating1-dehydro-6a-methyl-1l-ketotestosterone at room temperature in pyridinesolution with trimethylacetic anhydride results in1-dehydro-6a-methyl-1l-ketotestosterone 17-trimethylacetate, a lightcolored crystalline solid.

EXAMPLE 8 1 -dehydfo-6ot-m ethyl-1 1 -ket0test0ster0ne 17-cycl0-hexanecarboxy late In the same manner as shown in Example 3, treating1-dehydro-6ot-methyl-ll-ketotestosterone at room temperature in pyridinesolution with cyclohexanecarboxylic anhydride results in1-dehydro-6ot-methyl-1l-ketotestosterone 17-cyclohexanecarboxylate.

EXAMPLE 9 1 -dehydro-6a-methyl-1 1 -ket0test0ster0ne 17-phenylacetate Inthe same manner as shown in Example 3, treatingl-dehydro-6a-methyl-ll-ketotestosterone at room temperature in pyridinesolution with phenylacetic anhydride results in1-dehydro-6a-rnethy1-1l-ketotestosterone 17- phenylacetate.

EXAMPLE 10 1 -dehydr0-6a-methyl-11 -ket0test0sterone 1 7-phenylpropinate In the same manner as shown in Example 3, treating1-dehydro-6a-methyl-1l-ketotestosterone at room temperature in pyridinesolution with phenylpropionic anhydride results in1-dehydro-6u-methyl1l-ketotestosterone 17-phenylpropionate.

EXAMPLE 1 1 1 -dehydr0-6u-methyl1 1 -ket0test0sterone 1 7-acrylate Inthe same manner as shown in Example 3, treatingl-dehydro-6u-methyl-1l-ketotestosterone at room temperature in pyridinesolution with acrylic anhydride results inl-dehydro-6a-methyl-ll-ketotestosterone 17-acrylate.

EXAMPLE 12 1 -dehydr0-6u-methyl-1 1 -ketotestoster0ne 17-hemisuccinateIn the same manner as shown in Example 1, treating1-dehydro6m-methyl-1l-ketotestosterone at room temperature in pyridinesolution with succinic anhydride results in1-dehydro-6ez-methyl-1l-ketotestosterone 17-hemisuccinate, a lightcolored crystalline solid, which can be converted in aqueous alcoholicsolution to its sodium salt by the action of a sodium base such assodium carbonate.

In the same manner as shown in Examples 3 through 12, inclusive, other17-esters of 1-dehydro-6u-methyl-1lketotestosterone are prepared byreacting the corresponding acyl halides or acid anhydrides withl-dehydro-damethyl-1l-ketotestosterone. Esters thus prepared includethe, 17-butyrate, valerate, hexanoate, isobutyrate, isovalerate,cyclohexylacetate, adipate, crotonate, undecylenate, propiolate,undecolate, cinnamate ester of l-dehydro-6a-methyl-1l-fketotestosterone.

Using as. the stnrting material for the esterification the1-dehydro-6fi-methy1-1 l-ketotestosterone results in the corresponding 1dehydro-6B-methyl-1l-ketotestosterone l7-acylates, such as the17-acetate, propionate, benzoate, phenylacetate, phenylpropionate ofl-dehydro-GB-methyl- 1 l-ketotestosterone.

It is to be understood that the invention is not to be limited to theexact details of operation or exact compounds shown and described, asobvious modifications and equivalents will be apparent to one skilled inthe art, and the invention is therefore to be limited only by the scopeof the appended claims.

wherein R is selected from the substituents consisting of 25 2,783,226

keto oxygen wherein Ac is the acyl radical of a hydrocarbon carhoxylicacid containing from one to twelve carbon atoms, inclusive, and R isselected from the group consisting of L-HlethYl and 6fi-methyl.

2. l-dehydro-6u-methyladrenosterone.

3. 1-dehydro-6a-methyl-ll-ketotestosterone.

4. l-dehydro-Ga-rnethyl-ll-ketotestosterone, 17 -acylate wherein theacyl group is of a hydrocarbon carboxylic acid containing from one totwelve carbon atoms, inclusive.

S. 1-dehydro-6u-methy1- 11 -ketotestosterone 17-aceand state.

6. 1-dehydro-6u-methyl-1l-ketotestosterone 17-benzoate.

I 7. 1-dehydro-6a-methyl-1l-ketotestosterone 17 hemisuccinate and itssodium salt.

References Cited in the file of this patent UNITED STATES PATENTS GouldFeb. 26, 1957 OTHER REFERENCES Ringold et al.: J. Org. Chem., February1956, vol. 21,

so pages 239, 240.

UNITED STATES PATENT OFFICE CERTIFICATE OF {CORRECTION Jul 8,, 1958Patent No. 2,842,566 I Jean P. Rosselet It is hereby certified. thaterror appears in the-printed specification of the above numbered patentrequiring correction and that the said Letters Patent should read ascorrected below.

Column 5 lines 27 to 30, for

Signed and sealed this 19th day of *May 1959.

Attest:

KARL AXLINE 7 ROBERT C. WATSON v Commissioner of Patents AttestingOflicer

1. A COMPOUND OF THE FORMULA: